Toxin production and transcriptomic response to nitrate concentrations in the toxic dinoflagellate Alexandrium tamarense.

The bloom-forming dinoflagellate Alexandrium tamarense is one of the most important producers of paralytic shellfish poisoning toxins. Annually recurrent blooms of this dinoflagellate species is associated with the incremental nitrogen influx, especially excessive nitrate input. However, limited studies have been conducted on the toxin production and underlying molecular regulation mechanisms of A. tamarense under various nitrate (N) conditions. Therefore, toxin production and transcriptomic responses of this species were investigated. The toxin profile of A. tamarense was consistently dominated by the C2-toxins, and the cellular toxicity increased with N concentrations peaking at 9.23 ± 0.03 fmol/cell in the 883 µM N-added group. Under lower N conditions, expressions of two STX-core genes, sxtA and sxtG, were significantly down-regulated, suggesting that N regulated sxt expression and triggered responses related to toxin biosynthesis. Results of this study provided valuable insights into the ecophysiology of A. tamarense, enhancing our understanding of the occurrence of toxification events in natural environments.

Effects of sliver nanoparticles on nitrogen removal by the heterotrophic nitrification-aerobic denitrification bacteria Zobellella sp. B307 and their toxicity mechanisms.

Due to the widespread use of sliver nanoparticles (AgNPs), a large amount of AgNPs has inevitably been released into the environment, and there is growing concern about the toxicity of AgNPs to nitrogen-functional bacteria. In addition to traditional anaerobic denitrifying bacteria, heterotrophic nitrification-aerobic denitrification (HNAD) bacteria are also important participants in the nitrogen cycle. However, the mechanisms by which AgNPs influence HNAD bacteria have yet to be explicitly demonstrated. In this study, the inhibitory effects of different concentrations of AgNPs on a HNAD bacteria Zobellella sp. B307 were investigated, and the underlying mechanism was explored by analyzing the antioxidant system and the activities of key denitrifying enzymes. Results showed that AgNPs could inhibit the growth and the HNAD ability of Zobellella sp. B307. AgNPs could accumulate on the surface of bacterial cells and significantly destroyed the cell membrane integrity. Further studies demonstrated that the presence of high concentration of AgNPs could result in the overproduction of reactive oxygen species (ROS) and related oxidative stress in the cells. Furthermore, the catalytic activities of key denitrifying enzymes (nitrate reductase (NAR), nitrite reductase (NIR), and nitrous oxide reductase (N2OR)) were significantly suppressed under exposure to a high concentration of AgNPs (20 mg·L-1), which might be responsible for the inhibited nitrogen removal performance of strain B307. This work could improve our understanding of the inhibitory effect and underlying mechanism of AgNPs on HNAD bacteria.

Anisotropic mechanical properties of α-MoO3 nanosheets.

The mechanical behaviors of 2D materials are fundamentally important for their potential applications in various fields. α-Molybdenum trioxide (α-MoO3) crystals with unique electronic, optical, and electrochemical properties, have attracted extensive attention for their use in optoelectronic and energy conversion devices. From a mechanical viewpoint, however, there is limited information available on the mechanical properties of α-MoO3. Here, we developed a capillary force-assisted peeling method to directly transfer α-MoO3 nanosheets onto arbitrary substrates. Comparatively, we could effectively avoid surface contamination arising from the polymer-assisted transfer method. Furthermore, with the help of an in situ push-to-pull (PTP) device during SEM, we systematically investigated the tensile properties of α-MoO3. The measured Young's modulus and fracture strengths along the c-axis (91.7 ± 13.7 GPa and 2.1 ± 0.9 GPa, respectively) are much higher than those along the a-axis (55.9 ± 8.6 GPa and 0.8 ± 0.3 GPa, respectively). The in-plane mechanical anisotropy ratio can reach ∼1.64. Both Young's modulus and the fracture strength of MoO3 show apparent size dependence. Additionally, the multilayer α-MoO3 nanosheets exhibited brittle fracture with interplanar sliding due to poor van der Waals interaction. Our study provides some key points regarding the mechanical properties and fracture behavior of layered α-MoO3 nanosheets.

Comprehensive assessment of occurrence, temporal-spatial variations, and ecological risks of heavy metals in Jiaozhou Bay, China: A comprehensive study.

Heavy metals play a significant role in marine ecosystems, exerting notable impacts on the environment and human health. In this study, water, sediment, and aquatic organism samples from Jiaozhou Bay were investigated to comprehensively assess the distribution, temporal-spatial variations, and ecological risks of heavy metals. The results indicate that pollution from industrial wastewater discharge contributes to regional differences in the distribution of heavy metals, possibly being a major source of Zn, Cr, Cd, and Hg (r > 0.7, p < 0.05). Biological and physicochemical processes influence the distribution of Zn, Cr, and Pb in the water and sediment. Hg exhibits a polluted state in both the water and sediment, with As and Hg being the two highest-risk heavy metals in water and sediment, respectively. Among the organisms, crustaceans show significantly higher levels of heavy metal content and accumulation compared to mollusks and fish (p < 0.05), and the bioamplification of heavy metals occurs in the sediment-Rapana venosa-Portunus trituberculatus biological pathway. Portunus trituberculatus, Charybdis japonica, Oratosquilla oratoria, and Octopus ocellatus could pose risks to human health, especially for children and vulnerable populations. This study aims to enhance our understanding of the current status of heavy metal pollution in Jiaozhou Bay and to provide a scientific basis and favorable support for the ecological environmental protection and prevention of ecological risks associated with heavy metal pollution in Jiaozhou Bay and other bays in China.

Al8 Cluster-Based Metal Halide Frameworks: Balancing Singlet-Triplet Excited States to Achieve White Light and Multicolor Luminescence.

Luminescent metal clusters have attracted great interest in current research; however, the design synthesis of Al clusters with color-tunable luminescence remains challenging. Herein, an [Al8 (OH)8 (NA)16 ] (Al8 , HNA = nicotinic acid) molecular cluster with dual luminescence properties of fluorescence and room-temperature phosphorescence (RTP) is synthesized by choosing HNA ligand as phosphor. Its prompt photoluminescence (PL) spectrum exhibits approximately white light emission at room temperature. Considering that halogen atoms can be used to regulate the RTP property by balancing the singlet and triplet excitons, different CdX2 (X- = Cl- , Br- , I- ) are introduced into the reactive system of the Al8 cluster, and three new Al8 cluster-based metal-organic frameworks, {[Al8 Cd3 Cl5 (OH)8 (NA)17 H2 O]·2HNA}n (CdCl2 -Al8 ), {[Al8 Cd4 Br7 (OH)8 (NA)16 CH3 CN]·NA·HNA}n (CdBr2 -Al8 ) and {[Al8 Cd8 I16 (OH)8 (NA)16 ]}n (CdI2 -Al8 ) are successfully obtained. They realize the color tunability from blue to yellow at room temperature. The origination of fluorescence and phosphorescence has also been illustrated by structure-property analysis and theoretical calculation. This work provides new insights into the design of multicolor luminescent metal cluster-based materials and develops advanced photo-functional materials for multicolor display, anti-counterfeiting, and encryption applications.

Association of residential greenness, air pollution with adverse birth outcomes: Results from 61,762 mother­neonatal pairs in project ELEFANT (2011-2021).

BACKGROUND: Emerging evidence has demonstrated the benefits of greenness exposure on human health, while conflicts remain unsolved in issue of adverse birth outcomes. METHODS: Utilizing data from project ELEFANT spanning the years 2011 to 2021, we assessed residential greenness using the NDVI from MODIS data and residential PM2.5 exposure level from CHAP data. Our primary concerns were PTD, LBW, LGA, and SGA. Cox proportional hazard regression model was used to examine the association of residential greenness and air pollution exposure with risk of adverse birth outcomes. We performed mediation and modification effect analyses between greenness and air pollutant. RESULTS: We identified 61,762 mother­neonatal pairs in final analysis. For per 10 µg/m3 increase in PM2.5 concentration during entire pregnancy was associated with 19.8 % and 20.7 % increased risk of PTD and LGA. In contrast, we identified that an 0.1 unit increment in NDVI were associated with 24 %, 43 %, 26.5 %, and 39.5 % lower risk for PTD, LBW, LGA, and SGA, respectively. According to mediation analysis, NDVI mediated 7.70 % and 7.89 % of the associations between PM2.5 and PTD and LGA. Residential greenness could reduce the risk of PTD among mothers under 35 years old, living in rural areas, primigravidae and primiparity.. CONCLUSIONS: In summary, our results highlighted the potential of residential greenness to mitigate the risk of adverse birth outcomes, while also pointing to the adverse impact of PM2.5 on increased risk of multiple adverse birth outcomes (PTD and LGA). The significant mediation effect of NDVI emphasizes its potential as an important protective factor of PM2.5 exposure. Additionally, the identification of susceptible subgroups can inform targeted interventions to reduce adverse birth outcomes related to air pollution and lack of green spaces. Further research and understanding of these associations can contribute to better public health strategies aimed at promoting healthier pregnancies and birth outcomes.

Toward Clean 2D Materials and Devices: Recent Progress in Transfer and Cleaning Methods.

Two-dimensional (2D) materials have tremendous potential to revolutionize the field of electronics and photonics. Unlocking such potential, however, is hampered by the presence of contaminants that usually impede the performance of 2D materials in devices. This perspective provides an overview of recent efforts to develop clean 2D materials and devices. It begins by discussing conventional and recently developed wet and dry transfer techniques and their effectiveness in maintaining material "cleanliness". Multi-scale methodologies for assessing the cleanliness of 2D material surfaces and interfaces are then reviewed. Finally, recent advances in passive and active cleaning strategies are presented, including the unique self-cleaning mechanism, thermal annealing, and mechanical treatment that rely on self-cleaning in essence. The crucial role of interface wetting in these methods is emphasized, and it is hoped that this understanding can inspire further extension and innovation of efficient transfer and cleaning of 2D materials for practical applications.

Extra-High Mechanical and Phononic Anisotropy in Black Phosphorus Blisters.

Strain is an effective strategy to modulate the electrical, optical, and optoelectronic properties of 2D materials. Conventional circular blisters could generate a biaxial stretching of 2D membranes with notable strain gradients along the hoop direction. However, such a deformation mode cannot be utilized to investigate mechanical responses of in-plane anisotropic 2D materials, for example, black phosphorus (BP), due to its crystallographic orientation dependence. Here, a novel rectangular-shaped bulge device is developed to uniaxially stretch the membrane, and further provide a promising platform to detect orientation-dependent mechanical and optical properties of anisotropic 2D materials. Impressively, the derived anisotropic ratio of Young's modulus of BP flakes is much higher than the values obtained via the nanoindentation method. The extra-high strain-dependent phononic anisotropy in Raman modes along different crystalline orientations is also observed. The designed rectangular budge device expands the uniaxial deformation methods available, allowing to explore the mechanical, and strain-dependent physical properties of other anisotropic 2D materials more broadly.

Liquid Phase Edge Epitaxy of Transition-Metal Dichalcogenide Monolayers.

Precise monolayer epitaxy is important for two-dimensional (2D) semiconductors toward future electronics. Here, we report a new self-limited epitaxy approach, liquid phase edge epitaxy (LPEE), for precise-monolayer epitaxy of transition-metal dichalcogenides. In this method, the liquid solution contacts 2D grains only at the edges, which confines the epitaxy only at the grain edges and then precise monolayer epitaxy can be achieved. High-temperature in situ imaging of the epitaxy progress directly supports this edge-contact epitaxy mechanism. Typical transition-metal dichalcogenide monolayers (MX2, M = Mo, W, and Re; X = S or Se) have been obtained by LPEE with a proper choice of molten alkali halide solvents (AL, A = Li, Na, K, and Cs; L = Cl, Br, or I). Furthermore, alloying and magnetic-element doping have also been realized by taking advantage of the liquid phase epitaxy approach. This LPEE method provides a precise and highly versatile approach for 2D monolayer epitaxy and can revolutionize the growth of 2D materials toward electronic applications.

Effect of Ammonia on Laminar Combustion Characteristics of Methane-Air Flames at Elevated Pressures.

In this paper, laminar combustion characteristics of methane/ammonia/air flames are numerically investigated using the Chemkin/Premix code. The initial temperature is set as 298 K; the initial pressures are set as 1, 2, 5, 10, and 20 atm; and the equivalence ratios are set as 0.8-1.6. Laminar burning velocity (LBV); adiabatic flame temperature (AFT); net heat release rate (NHRR); and the mole fractions of H, NH2, NO, NO2, and HCN at stoichiometric ratio are studied with ammonia (NH3) addition. Meanwhile, temperature sensitivity and rate of production (ROP) are analyzed. The results show that with the increase of the initial pressures, LBV decreases and AFT and NHRR increase. With the increase of ammonia doping ratios, LBV, AFT, and NHRR decrease. From temperature sensitivity analyses, the main reactions that promote temperature rise are R39 (H + O2 < = > O + OH), R100 (OH + CH3 < = > CH2(S) + H2O), R102 (OH + CO < = > H + CO2), and R122 (HO2 + CH3 < = > OH + CH3O). The main reactions that inhibit temperature rise are R53 (H + CH3(+M) < = > CH4(+M)), R36 (H + O2 + H2O < = > HO2 + H2O), and R46 (H + HO2 < = > O2 + H2). For the rate of production of the free radical pool, the trends of H and NO are consuming first and then producing, and the trends of NH2, NO2, and HCN are the opposite. The pathway from methane to carbon dioxide is CH4 → CH3 → CH3O → CH2O → HCO → CO → CO2, and the pathway from ammonia to nitrogen is NH3 → NH2 → NH/HNO → NO/NO2 → N2.

Immobilization of Ochrobactrum sp. on Biochar/Clay Composite Particle: Optimization of Preparation and Performance for Nitrogen Removal.

The objective of this study was to prepare biochar/clay composite particle (BCCP) as carrier to immobilize Ochrobactrum sp. to degrade ammonia nitrogen (NH4 +-N), and the effects of calcined program and immobilizing material were investigated. Results reflected that the parameters were as follows: calcined temperature 400°C, heating rate 20°C min-1, and holding time 2 h, and the adsorption capacity could reach 0.492 mg g-1. Sodium alginate/polyvinyl alcohol, as embedding material, jointed with NH4 +-N adsorption process and then degraded by Ochrobactrum sp. with 79.39% degradation efficiency at 168 h. Immobilizing Ochrobactrum sp. could protect strain from high salt concentration to achieve the exceeding degradation efficiency than free bacteria, but could not block the impact of low temperature.

Toxicity of Naphthenic Acids on the Chlorophyll Fluorescence Parameters and Antioxidant Enzyme Activity of Heterosigma akashiwo.

Petroleum hydrocarbons can serve as a carbon source for marine phytoplankton; so, marine high-acid crude oil pollution events are likely to result in algal outbreaks or harmful algal blooms (HABs) in surface waters. Naphthenic acids (NAs) are the primary acidic component of crude oil, and red tide is of great concern due to its high diffusivity and strong destructive properties. It is important to study the mechanism of the toxic effect of NAs on the typical red tide algae, Heterosigma akashiwo, for the balance and stability of marine algae. The mechanism of NAs' damage effect was investigated in terms of the antioxidant enzyme activity, cell number, the chlorophyll positive fluorescence parameters, and the cell morphology of microalgae. Experiments confirmed the hormesis of low-concentration (0.5, 2, and 4 mg/L) NAs on Heterosigma akashiwo, and the indicators of high-concentration (8 and 16 mg/L) NA exposures showed inhibition. In this study, the toxic effect of NAs on the target organism showed a clear concentration-dose relationship. The 16 mg/L NAs stress caused severe damage to the morphology and structure of the target biological cells in a short time (96 h), and the population growth decreased. The target organisms showed a staged oxidative stress response to NAs. The behavior in the low-concentration treatment groups showed toxicant excitatory effects on the photosynthetic efficiency and antioxidant enzyme activity of the target organisms. This study provides theoretical and practical data for the development of an important toxicological model of the toxicant's excitement effects and antioxidant defense mechanisms. In addition, it provides prospective research data for the prediction and avoidance of ecological risk from NA pollution in marine environments.

A Comparative Study on Virology, Epidemiology, and Clinical Features of SARS and COVID-19.

In December 2019, an outbreak of an unknown cause of pneumonia (later named coronavirus disease 2019 [COVID-19]) occurred in Wuhan, China. This was found to be attributed to a novel coronavirus of zoonotic origin, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; previously named 2019 novel coronavirus or 2019-nCoV). The SARS-CoV-2, a new type of highly pathogenic human coronavirus related to severe acute respiratory syndrome coronavirus (SARS-CoV), spread rapidly worldwide and caused 246,303,023 confirmed infections, including 4,994,160 deaths, by October 31, 2021. SARS-CoV-2 and SARS-CoV vary in their specific characteristics, regarding epidemics and pathogenesis. This article focuses on the comparison of the virology, epidemiology, and clinical features of SARS-CoV and SARS-CoV-2 to reveal their common and distinct properties, to provide an up-to-date resource for the development of advanced systems and strategies to monitor and control future epidemics of highly pathogenic human coronaviruses.

Analysis of Emergency Medical Rescue in the "8-12" Tianjin Port Heavy Fire Explosion Accident.

With the progress in science and technology, hazardous chemicals are becoming more essential in chemical products, industrial and agricultural production, and daily life. Hazardous chemicals have poisoning, corrosive, explosive, and combusting natures; once on fire, they can trigger a chain of catastrophic incidences, resulting in casualties, property loss, and environmental pollution and posing hazards to life and property. Using the "8-12" explosion of the Ruihai Logistics warehouse in Tianjin Port (Binhai New District, China), the present study analyzes the characteristics of trauma of the casualties in this accident and the emergency medical rescue strategies. The goals were to improve the ability of emergency rescue in such accidents and to save people's lives and property to the maximum extent.

Characterization and complete genome analysis of Bacillus velezensis CB6 revealed ATP synthase subunit α against foodborne pathogens.

Given the serious threat of foodborne multidrug-resistant bacteria to animals and humans, finding an effective antibacterial compound has always been an important topic for scientists. Here, from the soil of Changbaishan, we have identified a bacterium that can inhibit the growth of Staphylococcus aureus. Nr genome database analysis and phylogenetic analysis showed that strain CB6 belongs to Bacillus velezensis. We found that the crude extract of strain CB6 has broad-spectrum antibacterial activity against foodborne pathogens. In addition, we showed that the crude extract loses antibacterial activity after treatment with papain. Next, strain CB6 was purified using ammonium sulfate precipitation, a Sephadex G-75 gel filtration column and high-performance liquid chromatography system (HPLC). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis indicated that the antibacterial compound was the protein ATP synthase subunit α (ATP-1), with a molecular weight of 55.397 KDa. Moreover, we reported the complete genome sequence of strain CB6, which is composed of a unique circular 3,963,507 bp chromosome with 3749 coding genes and a G + C content of 46.53%. The genome contained 12 gene clusters with antibacterial functions, which constituted over 20.947% of the complete genome. Of note, the amino acid sequence encoding the ATP-1 protein in the strain CB6 genome was identified. In addition to these findings, we speculate that the ATP-1 protein may provide energy for secondary metabolites, which in turn will improve the antibacterial activity of the secondary metabolites. All the above important features make the ATP-1 as a potential candidate for the development of new antibacterial drugs and food preservatives in the future.

Biological Characteristics and Genetic Analysis of a Highly Pathogenic Proteus Mirabilis Strain Isolated From Dogs in China.

To evaluate the antimicrobial resistance and virulence gene characteristics of highly pathogenic Proteus mirabilis. In this study, we isolated P. mirabilis CC15031 from diarrhea dogs in China, tested the median lethal dose (LD50), and measured the minimum inhibitory concentration (MIC) of 10 different antibiotics commonly used in veterinary clinic. Meanwhile, we presented the complete genome sequence annotations to analyze the virulence and resistance formation mechanism. The results showed that the CC15031 presented relatively potent pathogenicity in mice (LD50 = 0.57 × 106 CFU) and exhibited a high degree of resistance to all the tested antimicrobial agents. The CC15031 genome of 4,031,742 bp with 3,745 predicted genes had an average gene length of 917 bp and 38.99% guanine-cytosine content. A new variant of an integrative and conjugative element with a type IV secretion system (217,446 bp) conferring multidrug resistance was identified and characterized by structural analysis in CC15031. These data provide a foundation for understanding the genomic features and antimicrobial resistance mechanisms of this pathogen.

Genomic island type IV secretion system and transposons in genomic islands involved in antimicrobial resistance in Trueperella pyogenes.

Trueperella pyogenes (T. pyogenes) is a well-known opportunistic pathogen of many animal species. It can cause a variety of suppurative infections. The objective of this research was to get insight into the gene context and the location of the antimicrobial resistance determinants in the two multi-resistant T. pyogenes isolates TP3 and TP4. Comparative analysis of key factors leading to antimicrobial resistance was performed. Both isolates were resistant to erythromycin, azithromycin and tetracycline, and susceptible to ciprofloxacin, enrofloxacin, cefazolin and florfenicol. In addition, TP4 was resistant to amikacin and gentamicin. Whole-genome analyses revealed that both TP3 and TP4 contained two different genomic islands (TP3-GI1, TP3-GI5, TP4-GI5 and TP4-GI8) involved in multi-drug resistance. There is a common region in TP3-GI1 and TP4-GI5, containing the tetracycline resistance gene tet(W) and a series of genes involved in type IV secretion systems. Several genes located on TP3-GI5 and TP4-GI8 are highly homologous. Tetracycline-resistance gene tet(33) was potentially acquired by horizontal gene transfer via IS6100 located on 57,936 bp TP3-GI5. The macrolide resistance gene erm(X) was located near the end of the TP3-GI5. The sequence analysis of TP4-GI8 showed that two copies of erm(X) and two IS1634 elements located in the same orientation may have formed a composite transposon. GI-type T4SS, transposons and multiple resistance genes located on GIs play a key role in multiple drug resistance of TP3 and TP4.

Ulinastatin ameliorates acute kidney injury induced by crush syndrome inflammation by modulating Th17/Treg cells.

BACKGROUND: Acute kidney injury (AKI) is the main complication of crush syndrome (CS), and it is also a cause of lethality in CS. However, effective treatments for AKI are still lacking. Ulinastatin (UTI) is a broad-spectrum serine protease inhibitor extracted from human urine that reportedly modulates innate immunity and pro-inflammatory responses in sepsis. Here, we explored the effect and the potential mechanism of ulinastatin on crush syndrome-induced acute kidney injury (CSAKI). METHODS: A CSAKI rat model was established by using a digital crush injury device platform. Forty-six male Wistar rats were randomly divided into five groups: the normal control (n = 6), CSAKI model (n = 10), CSAKI plus UTI1 (50,000 U/kg) (n = 10), CSAKI plus UTI2 (100,000 U/kg) (n = 10) and CSAKI plus UTI3 (200,000 U/kg) (n = 10) groups. Hematoxylin-eosin (HE) staining was used to investigate the reliability of the CSAKI model. The percentage of Th17/Treg lymphocytes in peripheral blood was measured by flow cytometry, and the expression of transcription factors associated with Th17/Treg cells was evaluated by quantitative real-time polymerase chain reaction (PCR). In addition, specific cytokines released by Th17/Treg cells in serum and kidney tissues were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: Treatment with ulinastatin could significantly decrease serum BUN, CK, Scr, Mb and K+ levels compared with CSAKI group. HE staining results showed that ulinastatin could inhibit inflammatory cells infiltration, decrease sarcomere rupture in muscle tissues induced by extrusion, and alleviate the glomerular congestion and edema, as well as decrease myoglobin cast in kidney tissues. The proportion of CD4+CD25+Foxp3+ regulatory T (Treg) cells and Foxp3 expression levels were decreased in the CSAKI animals, while IL-17 expression levels were significantly increased, compared with those of the normal control group. Treatment with ulinastatin upregulated the proportion of Treg cells in CD4+ T cells and downregulated the expression of IL-17 compared with those of the CSAKI group. CONCLUSION: The findings of our study indicate that UTI attenuates CS-induced AKI and alleviate the inflammatory response during the early stage. The mechanism of UTI may be due to regulating the balance between Th17/Treg cells. Our study provides a new mechanism for the beneficial effect of ulinastatin on CSAKI.

Multidrug resistance genes are associated with a 42-kb island TGI1 carrying a complex class 1 integron in Trueperella pyogenes.

OBJECTIVES: This research was conducted to ascertain the context and location of the antibiotic resistance determinants in a multiple antibiotic-resistant Trueperella pyogenes isolate TP1. METHODS: The genome was sequenced using PacBio RS II, and the filtered data were assembled using Canu. Sequences were annotated on the basis of those in GenBank, and the genomic island (GI) of the TP1 was predicted by IslandPath-DIMOB. RESULTS: TP1 as a multiple antibiotic-resistant isolate was recovered at Jilin Province (China) in 2017 from a dairy cow with pneumonia. TP1 exhibited resistance to aminoglycosides (gentamicin and amikacin), macrolides (erythromycin), lincosamides (clindamycin), sulfonamides (sulfamonomethoxine), tetracyclines (tetracycline and doxycycline) and chloramphenicols (chloramphenicol and florfenicol). An antibiotic resistance gene clustered together with the aadB, aadA1, cmlA5 and cmlA6 resistance genes located on a 7-kilobase (kb) multidrug-resistant (MDR) region, constituting a complex class 1 integron. The MDR region was located at one end of a 42-kb GI, and IS6100Δ1 mediated a genetic rearrangement with the complex class 1 integron-like SGI1 and formed a composite transposon. Furthermore, the tetW gene was located outside the four GIs consistent with tetracycline and doxycycline resistance. The ermD gene positioned in the front end of the 42-kb GI played an important role in mediating acquired erythromycin and clindamycin resistance. CONCLUSIONS: Multiple resistance genes are located in a complex class 1 integron within a 42-kb T. pyogenes genomic island (TGI1), leading to TP1 multiple drug resistance. In comparison with SG1 families, TGI1 possesses versatile gene distribution and specific gene context for it upstream and downstream, and it represents a new lineage of genomic resistance islands.

Transcriptome Analysis Reveals AI-2 Relevant Genes of Multi-Drug Resistant Klebsiella pneumoniae in Response to Eugenol at Sub-MIC.

Eugenol, the major active essential oil component of clove, was reported to possess QS (quorum sensing) inhibitory activity. A previous study found that eugenol could bind to quorum sensing receptors of Pseudomonas aeruginosa and down-regulate the expression of Streptococcus mutans virulence genes at sub-MIC (minimum inhibitory concentration) without affecting the bacterial growth. However, the alterations of QS signal molecules at transcription levels was not well understood. To better understand interactions of Klebsiella pneumoniae in response to eugenol and explore molecular regulations, transcriptome sequencing was performed. A total of 5779 differentially expressed genes (DEGs) enriched in a variety of biological processes and pathways were identified. The transcriptional data was validated by qPCR and the results showed that the expression profiles of 4 major genes involved in autoinducers-2 (AI-2) synthesis, including luxS, pfs, and lsrK were consistent with transcriptome analysis except for lsrR, a transcriptional repressor gene of lsr operon, which may repress the expression of following genes responsible for AI-2 signal transmission in vivo. In vitro AI-2 synthesis assay also revealed that eugenol could inhibit AI-2 generation. The results of our study offer insights into the mechanisms of QS inhibitory activity and K. pneumoniae AI-2 alterations after eugenol treatment.